| 质粒名称: | pQCascade-tra13 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 140628 |
| 存储实验室: | Sheng Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pCDFDuet-1 |
| 完整质粒大小: | 8510bp |
| 载体类型: | Bacterial Expression, CRISPR |
| 载体抗性: | Streptomycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | VchTniQ, VchCas8, VchCas7, VchCas6, CRISPR-tra13 |
| 物种来源: | Vibrio cholera |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Unknown (unknown if destroyed) |
| 3’克隆位点: | Unknown (unknown if destroyed) |
|
文献引用方法 |
|
| 材料和方法部分: | pQCascade-tra13 was a gift from Sheng Yang (Addgene plasmid # 140628 ; http://n2t.net/addgene:140628 ; RRID:Addgene_140628) |
| 参考文献部分: |
Multicopy chromosomal integration using CRISPR-associated transposases. Zhang Y, Sun X, Wang Q, Xu J, Dong F, Yang S, Yang J, Zhang Z, Qian Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S. ACS Synth Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073. 10.1021/acssynbio.0c00073 PubMed 32551502
|
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pQCascade-tra13 8510 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Expresses V. cholerae TniQ, Cas8, Cas7, and Cas6 and crRNA-tra13.
The crRNA-tra13 targets 13 copies transposon loci in Tatumella
citrea..
ACCESSION .
VERSION .
KEYWORDS pQCascade-tra13
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8510)
AUTHORS Zhang Y, Sun X, Wang Q, Xu J, Dong F, Yang S, Yang J, Zhang Z, Qian
Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S
TITLE Multicopy chromosomal integration using CRISPR-associated
transposases.
JOURNAL ACS Synth Biol. 2020 Jun 17. doi: 10.1021/acssynbio.0c00073.
PUBMED 32551502
REFERENCE 2 (bases 1 to 8510)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..8510
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 73..91
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 92..116
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind complement(4914..4932)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 4928..4975
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
CDS complement(5146..5937)
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
promoter complement(5938..6029)
/gene="bla"
/label=AmpR promoter
CDS complement(7090..8172)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=lacI
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 8134..8153
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(8173..8250)
/gene="lacI"
/label=lacI promoter
promoter 8296..8314
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 8315..8339
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
ORIGIN
1 caagcttgcg gccgcataat gcttaagtcg aacagaaagt aatcgtattg tacacggccg
61 cataatcgaa attaatacga ctcactatag gggaattgtg agcggataac aattccccat
121 cttagtatat tagttaagta taagaaggag atatacatat gtttttgcaa agacctaaac
181 cttacagcga tgaaagttta gaaagtttct ttatccgagt ggctaacaaa aatggctacg
241 gtgatgtcca tcgcttccta gaagccacta aacgattcct tcaagacatt gaccataatg
301 gctatcaaac ctttccgact gatataactc ggataaaccc atactcagct aaaaacagtt
361 ccagcgcacg aactgcgtca ttcctgaaac ttgcacaatt gacatttaat gaaccgccag
421 agctacttgg gttggcaatt aacagaacaa acatgaaata ctcgccgtca actagcgcgg
481 ttgttagagg tgcagaagtc tttcctcgca gtttactacg gacgcactcc atcccctgct
541 gtcctttgtg tctgcgagaa aatggctacg cctcctacct ttggcacttt caggggtacg
601 aatactgcca cagccataac gtacctttaa ttaccacttg tagctgtggt aaggagtttg
661 actaccgagt atctgggtta aagggcattt gctgcaaatg caaggagcct atcaccttaa
721 ccagcaggga gaacggtcat gaggcagcgt gtactgtttc aaactggctt gctggccatg
781 aatctaaacc tctgccaaat cttcctaaaa gctaccgatg gggtttagtt cattggtgga
841 tgggtattaa agatagcgag ttcgatcact tttcgttcgt tcaatttttc tcaaactggc
901 caaggtcatt ccactcgata atcgaagatg aagtagagtt caaccttgag catgctgttg
961 tcagcacgtc tgaattacga ctaaaagatc ttcttggtcg attgtttttc ggttcaattc
1021 ggttacctga gcggaatctt caacacaata tcatccttgg tgagcttctc tgctatttag
1081 aaaatcgttt atggcaagac aagggattaa tcgccaacct caaaatgaac gcgttagagg
1141 cgactgtaat gttaaattgt agcctcgatc agattgcatc aatggttgaa caacgcatct
1201 tgaagccaaa tcgaaaaagc aagcccaaca gccctcttga tgttaccgat tatctatttc
1261 atttcggcga tattttctgt ctttggttag ctgagttcca aagcgatgag tttaaccgtt
1321 cgttttatgt gtcgaggtgg taaatgcaaa ctctgaaaga actaatcgca tccaatcctg
1381 acgacttaac aactgaactt aagagagcat ttcgtccact cacaccgcat attgcaattg
1441 atggtaatga acttgacgca ctgacgatat tagtcaattt aaccgataag actgatgatc
1501 agaaagacct gctcgatcga gccaaatgca agcaaaaact tcgagatgaa aaatggtggg
1561 ctagctgcat aaattgcgtt aattacagac aaagccataa cccaaaattc ccggatatac
1621 gttctgaagg cgtgattcga acccaagccc tgggtgaatt accgagtttc ctactctcgt
1681 cctcaaaaat cccaccatac cattggtcat atagccatga ttcaaaatac gtcaacaaaa
1741 gcgcattcct caccaatgag ttttgttggg atggtgagat ctcatgtttg ggtgagcttc
1801 ttaaagatgc agatcaccca ctttggaata ctttgaaaaa gttaggttgt tctcaaaaaa
1861 cttgcaaagc aatggcaaaa caactagctg atattactct cacgactatc aatgtcacac
1921 tcgcaccaaa ttacctgact caaatatctc tccccgatag tgatacatcc tacatttcac
1981 tctctcctgt tgcatcgcta tcgatgcaaa gccactttca tcagaggctc caagatgaaa
2041 atcggcatag cgcgataacg cggtttagcc gaactaccaa catgggggtc actgcgatga
2101 catgtggcgg cgcatttaga atgttaaagt ctggcgctaa gttttccagc ccccctcatc
2161 accgattaaa cagtaaacga agttggttga cgtcagagca tgttcagtca ttaaaacagt
2221 accagcgcct caacaaaagc ctcatacctg aaaactctcg gattgcactc cgtagaaaat
2281 acaaaatcga gcttcaaaat atggtcagat cttggtttgc aatgcaagac catacccttg
2341 attcgaacat acttatccaa catttgaatc atgacctatc ttacttagga gccacaaaac
2401 gttttgcata cgatccagcg atgaccaagc tctttactga gcttttgaaa cgagagttat
2461 caaattcaat caataatggt gagcaacaca ctaatggatc gtttttagtc ctaccgaata
2521 tcagggtttg tggcgcaaca gctttaagct ccccggtaac ggtggggatt ccatcactta
2581 cagctttctt tggcttcgtt cacgcatttg aacggaatat aaatcgcacc acctcatcgt
2641 ttcgtgttga atcctttgcg atatgcgtcc atcaactaca tgtcgagaag cgaggtttga
2701 cagcagagtt tgtggaaaaa ggcgacggga ctatatccgc tcccgcgacc cgggatgact
2761 ggcagtgtga tgtcgtattt agccttattt tgaacaccaa ctttgctcaa catattgacc
2821 aagatacgtt agttacatca ctaccaaagc gattggctcg gggttcagca aaaattgcga
2881 ttgatgactt taaacatatc aactcattct cgacattaga aacagcgatc gaatctctgc
2941 caatagaagc tggtaggtgg ttatcacttt acgcacagtc aaacaataat ctaagtgatc
3001 tattagcagc catgacagag gaccatcagc tcatggcaag ctgcgtcggt taccacttgt
3061 tagaagagcc caaagataaa ccaaactccc tcagaggtta caaacacgct atcgccgagt
3121 gcatcattgg actcattaac tcaatcacct ttagctcaga gactgatccc aacacaatct
3181 tttggtcgct aaagaactat caaaactacc tagtggtaca gccaaggagt atcaacgatg
3241 aaactaccga caaatctagc ctatgagcgc tctatcgacc catcagatgt ctgttttttt
3301 gtcgtctggc ccgatgatag aaaaacacct ttaacctaca attctcgtac tctgctcggg
3361 caaatggaag cggcatcatt agcctatgat gtctcaggtc aaccaataaa aagtgccacc
3421 gctgaggcgt tagctcaagg gaaccctcat caagttgatt tctgccacgt tccatacggt
3481 gcgagtcata ttgaatgcag tttctccgtc tcgttttctt ctgaactacg tcaaccatat
3541 aagtgtaact caagcaaagt taaacaaacg ctagtgcaat tagtcgagct ctacgaaacg
3601 aaaatcggct ggactgagct agcaacccga tatttgatga acatttgcaa cggtaaatgg
3661 ctgtggaaaa atacccgtaa agcctattgc tggaacattg tacttacacc ttggccttgg
3721 aacggggaaa aggttggatt tgaagatatt cgtactaact acacctcacg gcaagacttt
3781 aaaaataata aaaattggtc tgctatagtt gaaatgatca aaaccgcatt ttctagtact
3841 gatgggctgg cgatatttga agtcagggcc accttgcact tgccaacgaa tgctatggtg
3901 cggccaagcc aagttttcac agaaaaagaa agtggcagta aaagtaaatc taaaactcaa
3961 aacagtcgag tttttcagag tacaactatt gatggtgaac gatcgccaat actaggggcc
4021 tttaaaacgg gagcagctat tgcaaccatt gacgactggt atcctgaagc cactgagcca
4081 ctaagggtcg gacggtttgg ggttcatcgc gaagatgtca cttgctaccg tcatccgtct
4141 accggaaaag attttttctc gatattacaa caagcagagc actatattga agtgttgagc
4201 gccaacaaaa ctcccgctca agaaactatc aacgacatgc actttttaat ggctaacctg
4261 attaagggtg ggatgttcca gcataaagga gactgactgt gaaatggtat tataagacaa
4321 tcacctttct gccagagttg tgcaacaacg agtcactggc tgcaaagtgt ctccgcgttc
4381 tgcatggatt taactatcag tatgagacac gaaatatcgg cgtttcattt ccgctttggt
4441 gtgatgcaac ggttggaaaa aagatttcat ttgtcagcaa gaacaagata gaactcgact
4501 tactacttaa acaacactat ttcgtccaaa tggaacaact tcaatacttt catatatcca
4561 acactgttct cgtcccagaa gattgtacat acgtttcctt tagacgctgt caatctatag
4621 ataagctcac agcagcaggg ctggcaagga aaatcagacg cctggagaaa cgtgctctat
4681 ctagaggcga gcaatttgac ccatcatctt ttgctcaaaa agagcatact gcaatagcgc
4741 actaccactc acttggggag tccagcaaac agacgaaccg caactttcga ctcaatatca
4801 ggatgctctc ggagcaaccc cgtgagggga actcgatttt tagtagctat ggcttatcaa
4861 attcagaaaa ctcgtttcag cctgtaccct taatctgacc taggctgctg ccaccgctga
4921 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
4981 acctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
5041 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcatc
5101 gtggccggat cttgcggccc ctcggcttga acgaattgtt agacattatt tgccgactac
5161 cttggtgatc tcgcctttca cgtagtggac aaattcttcc aactgatctg cgcgcgaggc
5221 caagcgatct tcttcttgtc caagataagc ctgtctagct tcaagtatga cgggctgata
5281 ctgggccggc aggcgctcca ttgcccagtc ggcagcgaca tccttcggcg cgattttgcc
5341 ggttactgcg ctgtaccaaa tgcgggacaa cgtaagcact acatttcgct catcgccagc
5401 ccagtcgggc ggcgagttcc atagcgttaa ggtttcattt agcgcctcaa atagatcctg
5461 ttcaggaacc ggatcaaaga gttcctccgc cgctggacct accaaggcaa cgctatgttc
5521 tcttgctttt gtcagcaaga tagccagatc aatgtcgatc gtggctggct cgaagatacc
5581 tgcaagaatg tcattgcgct gccattctcc aaattgcagt tcgcgcttag ctggataacg
5641 ccacggaatg atgtcgtcgt gcacaacaat ggtgacttct acagcgcgga gaatctcgct
5701 ctctccaggg gaagccgaag tttccaaaag gtcgttgatc aaagctcgcc gcgttgtttc
5761 atcaagcctt acggtcaccg taaccagcaa atcaatatca ctgtgtggct tcaggccgcc
5821 atccactgcg gagccgtaca aatgtacggc cagcaacgtc ggttcgagat ggcgctcgat
5881 gacgccaact acctctgata gttgagtcga tacttcggcg atcaccgctt ccctcatact
5941 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
6001 atttgaatgt atttagaaaa ataaacaaat agctagctca ctcggtcgct acgctccggg
6061 cgtgagactg cggcgggcgc tgcggacaca tacaaagtta cccacagatt ccgtggataa
6121 gcaggggact aacatgtgag gcaaaacagc agggccgcgc cggtggcgtt tttccatagg
6181 ctccgccctc ctgccagagt tcacataaac agacgctttt ccggtgcatc tgtgggagcc
6241 gtgaggctca accatgaatc tgacagtacg ggcgaaaccc gacaggactt aaagatcccc
6301 accgtttccg gcgggtcgct ccctcttgcg ctctcctgtt ccgaccctgc cgtttaccgg
6361 atacctgttc cgcctttctc ccttacggga agtgtggcgc tttctcatag ctcacacact
6421 ggtatctcgg ctcggtgtag gtcgttcgct ccaagctggg ctgtaagcaa gaactccccg
6481 ttcagcccga ctgctgcgcc ttatccggta actgttcact tgagtccaac ccggaaaagc
6541 acggtaaaac gccactggca gcagccattg gtaactggga gttcgcagag gatttgttta
6601 gctaaacacg cggttgctct tgaagtgtgc gccaaagtcc ggctacactg gaaggacaga
6661 tttggttgct gtgctctgcg aaagccagtt accacggtta agcagttccc caactgactt
6721 aaccttcgat caaaccacct ccccaggtgg ttttttcgaa ggttaagtca gttggggaac
6781 tgcttaacct tcgatcaaac cacctcccca ggtggttttt tcgtttacag ggcaaaagat
6841 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctactg aaccgctcta
6901 gatttcagtg caatttatct cttcaaatgt agcacctgaa gtcagcccca tacgatataa
6961 gttgtaattc tcatgttagt catgccccgc gcccaccgga aggagctgac tgggttgaag
7021 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
7081 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
7141 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
7201 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
7261 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
7321 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tgtccgcacc
7381 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
7441 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
7501 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
7561 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
7621 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
7681 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
7741 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
7801 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
7861 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
7921 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
7981 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
8041 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
8101 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
8161 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
8221 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
8281 cctgcattag gaaattaata cgactcacta taggggaatt gtgagcggat aacaattccc
8341 ctgtagaaat aattttgttt aactttaata aggagatata ccatgggtga actgccgagt
8401 aggtagctga taactgaaaa tgcagaggcc ttcggcaaag catctagtga actgccgagt
8461 aggtagctga taacggatcc gaattcgagc tcggcgcgcc tgcaggtcga
//
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